Aryl esters of {60 -carboxyarylmethyl-penicillins

ABSTRACT

Arylchloro (and bromo) carbonyl ketenes, arylcarboxy ketene aryl esters derived therefrom, methods for their preparation and the use of the esters as acylating agents for the production of antibacterial agents; namely, aryl esters of Alpha carboxyarylacetyl derivatives of 6-aminopenicillanic acid, and by hydrolysis, the corresponding acid derivatives, are described.

United States Patent Butler [4 July 25, 1972 [54] ARYL ESTERS 0F 0!- I [56] References Cited ARBOXY YLMEIHYL-PE C AR NICILLINS UNITED STATES PATENTS 72 I t K th 13 tle Old L C I 1 em yme 3,502,656 3/1970 Neal et al ..260/239.1 [73] Assignee: Pfizer Inc., New York, NY.

a Primary ExaminerNicholas S. Rizzo [22] Flled' May 1970 Attomey-Connolly & Hut: [211 Appl. No.: 38,903

[57] ABSTRACT Related [1.8. Application Data Arylchloro (and bromo) carbonyl ketenes, arylcarboxy commuauonm'pal't of 330516, June 4, ketene aryl esters derived therefrom, methods for their 1969' abandoned, Cmtmuauondmpan of preparation and the use of the esters as acylating agents for Jan 1968, abandonedthe production of antibacterial agents; namely, aryl esters of a-carboxyarylacetyl derivatives of 6-aminopenicillanic acid, [52] U.S.CI ..424/27l,260/239.l and by hydrolysis the corresponding i derivatives, are 511 Im. CI. l ..C07d 99/16 described [58] Field ofSearch ..260/239.1;424/271 8 Claims, No Drawings ARYL. ESTERS OF a-CARBOXYARYLMETHYL- PENICILLINS CROSS-REFERENCE TO RELATED APPLICATIONS BACKGROUND OF THE INVENTION This invention relates to a series of novel antibacterial agents; namely, esters of a-carboxyarylmethylpenicillins, to the pharmaceutically-acceptable salts thereof, and to methods for their preparation.

The production of ketenes from malonic acid derivatives is described in the literature: Staudinger, Helv. Chim. Acta 8, 306 1925 prepared a series of low-molecular-weight dialkyl ketenes by the thermal decomposition of di(lwer)alkyl substituted malonic anhydrides; Staudinger et al., Ibid, 6, 291 (1923) and Ber. 46, 3539 (1913) produced various disubstituted ketenes by thermal decomposition of mixed anhydrides prepared from disubstituted malonic acids and diphenyl ketenes. A further method comprises the dehalogenation of a-halo acyl halides with zinc (Staudinger, Ann. 356, 71 (1907); 380, 298 (1911)). By extension ofthis reaction, Staudinger et al., Ber. 42, 4908 (1909) prepared ethyl carbethoxy ketene by the dehalogenation of diethyl-abromo-a-ethyl malonate. Another method, the decomposition of diazo ketones, has been used to prepare certain diaryl ketenes (Smith et al., Org. Syntheses 20, 1940; Gilman et al., Rec. trav. chim. 48, 464, 1929). It is further known that certain disubstituted acetyl chlorides undergo dehydrohalogenation under the influence of tertiary amines to form keto ketenes. This method, however, appears to be limited to the preparation of certain aryl and high-molecular-weight keto ketenes, all of which are relatively resistant to dimerization (Staudinger et al., Ber. 41, 594, I908).

The reaction of phenylmalonic acid with'phosphorus pentachloride (1:2 molar ratio) in ether solution is reported by Sorm et al. (Collection Czechoslov. Chem. Communs. 20, 593-6, 1955) to produce phenylmalonyl chloride. The same authors report (loc. cit.) that when the reaction is conducted in' the absence of a solvent at the reflux temperature, phenylchloromalonyl chloride is produced. Both products are isolated by vacuum distillation.

The preparation of lower alkyl esters-of phenylcarboxy ketene by thermal decomposition of diazoketo esters has been described by Staudinger et al. (Ber. 49, 2522, 1916). However, the method used by Staudinger is rather complex and results, on an over-all basis, in rather pooryields. The process of the present invention for making such esters, on the other hand, is simple and productive of satisfactory yields.

The preparation of (a,a-dimethylbenzyl)carbethoxyketene by dehydrochlorination, pyrolytically or with triethylamine, of a-carbethoxy-Bphenyl-isovaleryl chloride which is prepared by the reaction of thionyl chloride and a-carbethoxy-B-phenylisovaleric acid is reported by Newman et al., J.'Org. Chem. 27, 1436-8 (1962). Also reported by these same investigators is t-butylcarbethoxyketene. This latter compound could not be prepared by pyrolytic dehydrochlorination but required the use of triethylamine. Both a-carbethoxyketenes are resistant to dimerization by virtue of the bulkiness of the a,a-dimethylbenzyl and the t-butyl groups.

The use of ketenes as acetylating agents is well-known in the art. The acylation of amino groups by means of simple or mixed acid anhydrides, acid halides, acid azides, B- thiolacetones, acylated enols and carboxylic acids with carbodiimides is also known in the art (Sheenan, US. Pat. No. 3,159,617, Dec. 1, 1964). French Pat. 1,427,686, Jan. 3, 1966, describes the acylation of 6-aminopenicillanic acid with ketenes, including acylation with (a,a-dimethylbenzyl)carbethoxyketene to produce potassium 6-(N-carbethoxyphenylisovaleryl)aminopenicillanate; more conveniently referred wherein R broadly referred to herein as an aryl group, is

to as potassium a-carbethoxy-fi,[3-dimethyl-B-phenethylpenicillin. However, the preparation of a-carboxyarylmethyh penicillins has, up until now, been limited to the use of a simple or mixed anhydride, an acid halide of an arylmalonic acid or an arylmalonic acid ester as acylating agent (U.S. Pat. No. 3,142,673, British Pat. No. 1,004,670).

SUMMARY OF THE INVENTION It has now been unexpectedly found that a variety of arylchlorocarbonyl ketenes, and the corresponding bromo analogs, can readily be prepared by the reaction of arylmalonic acids with a halogenating agent followed by thermal elimination of hydrogen halide and vacuum distillation of the reaction product thus formed. The process and the compounds produced are summarized by the reaction:

COOH

selected from the .group consisting of: thienyl, furyl, pyridyl, phenyl, and substituted phenyl wherein the substituent is selected from the group consisting of (lower)alkyl, chloro, bromo, (lower)alkoxy, di(lower)alkylamino and trifluoromethyl, and X is selected from the group consisting of chloro and bromo.

The process of this invention, in view of the teaching of Sormet al. (loc. cit.) that phenylmalonyl chloride and phenylchloromalonyl chloride are obtained by the action of phosphorous pentachloride on phenylmalonic acid in the presence of a solvent and the products recovered by vacuum distillation,'is most surprising and unexpected. Repetition of the Sorm et al. procedure for making phenylmalonyl chloride has been found to produce phenylchlorocarbonyl ketene rather than phenylmalonyl chloride. The existence of the ketene compound is completely unrecognized by Sorm et al.

The process, in general, comprises reacting an aryl substituted malonic acid with a halogenating agent selected from the group consisting of P(X) P(X) PO(X) and SO(X) wherein X is as defined above at a temperature of from about 0 C. to about 50 C. The dihalide thus produced is thermally decomposed at about to C. to provide the aryl halocarbonyl ketene.

The arylhalocarbonyl ketenes exhibit a dual functionality and react both as acid halides and ketenes. They are, therefore, valuable as intermediates for further synthesis. Alcohols (R Ol-I), for example, react with the aryl halocarbonyl ketenes at low temperatures, e. g., from about 70 C. to about 30 C. to produce the corresponding esters of aryl carboxy ketenes which are useful as acylating agents. Reaction appears to occur first with the ketene group to form a transient intermediate which rearranges with elimination of hydrogen halide to the arylcarboxy ketene ester.

The novel arylcarboxy ketene esters of this invention are especially valuable as agents for the acylation of amines with production of esters of a-carboxy-a-arylacetylamines. They are particularly useful for the acylation of amines such as 6- aminopenicillanic acid for the production of known and novel a-carboxyarylmethylpenicillins useful as antibacterial agents. Prior art methods for introducing a-carboxyarylacetyl groups into amino compounds such as 6-aminopenicillanic acid to produce a-carboxyarylmethylpenicillins have made use of acid anhydrides, mixed or simple, or acid halides of arylmalonic acids. The use of the prior acylating agents requires extreme caution during reaction and recovery steps in order to obtain satisfactory yields of these relatively unstable penicillins which are susceptible to decarboxylation of the carboxy group. The acylating agents of this invention, on the other hand, react smoothly and rapidly with amines at low temperatures and produce no undesirable by-products.

The acid halides and esters of the aryl carboxy ketenes form 3-aryl substituted 2-oxo-oxetenes (l,3-epoxypropenes) in solution. These compounds react in a manner analogous to that of the compounds from which they are derived, and for purposes of this invention are equivalent to said compounds.

The above reactions are summarized in the following sequence:

wherein Y; is selected from the group consisting of:

wherein Z is lower alkylene and is selected from the group consisting of (CH and -(CH and substituted derivatives thereof wherein the substituent is selected from the group consisting of methyl, chloro, and bromo; and R is:

: H-COOH Of the lower alkyl, lower alkoxy and lower alkanoyl groups those having from I to 4 carbon atoms in the alkyl, allcoxy and alkanoyl moieties are preferred since the reactants bearing such groups are more readily available than those required for such groups showing a greater number of carbon atoms.

Also included within the scope of this invention are the pharmaceutically-acceptable salts of the novel compounds of formula lll in which one or both acid groups are involved in salt formation. Salts such as the sodium, potassium, calcium. magnesium, ammonium and substituted ammonium salts, e.g., procaine, dibenzylamine, N,N'-dibenzylethylenediamine, N,N'-bis-(dehydroabietyl)ethylenediamine, l-ephenamine, N-methyl morpholine, N-methylpiperidine, N-ethylpiperidine, N-benzyl-B-phenethylamine, trialkylamines, including triethylamine, as well as salts with other amines which have been used to form salts with benzylpenicillin are useful for the preparation of pharmaceutically-elegant compositions of these valuable antibiotics.

DETAILED DESCRIPTION OF THE INVENTION The production of ar'ylchloro (and bromo) carbonyl ketenes comprises the reaction of an arylmalonic acid with a halogenating agent selected from the group consisting of P(X) P(X) F000;, and SO(X) wherein X is as defined above at temperatures ranging from about 0 C. to about 50 C. for periods ranging from about 1 hour to about 10 hours. The reaction is conducted in the presence of a solvent system, preferably a reaction inert solvent system. Suitable solvents are dialkyl ethers, e.g., diethyl ether, dipropyl ether, monoand dimethyl ethers of ethylene glycol and propylene glycol, methylene chloride and chloroform.

The reaction period, of course, is dependent upon the reaction temperature and the nature of the reactants. However, for a given combination of reactants, the lower temperatures require longer reaction periods than do higher temperatures.

The molar proportions of reactants, i.e., arylmalonic acid and halogenating agent, can vary widely, e.g., up to 1:10 or higher, but for satisfactory yields should be at least stoichiometric. In actual practice the stoichiometric ratio of reactants is preferred.

The reactants may be added all at once or separately. If separately, the order of addition is not critical. However, it appears that the reaction is smoother and subject to fewer sidereactions, as evidenced by the color of the reaction mixture, particularly upon distillation, when the arylmalonic acid is added to the halogenating agent. The reaction mixture, under such conditions, generally progresses from a yellow to a red color. The reaction mixture on reverse addition, i.e., the addition of halogenating agent to the arylmalonic acid, progresses from yellow to black.

The arylhalocarbonyl ketene products are isolated from the reaction by distillation in vacuo. Because of their great reactivity they are generally stored under a nitrogen atmosphere at low temperatures and in the absence of light.

Reaction of the arylhalocarbonyl ketenes with alcohols is conducted on a 1:1 molar ratio at a temperature of from about 70 C. to about 30 C. when conversion of the arylhalocarbonyl ketene to a ketene ester is desired. A reaction-inert solvent, such as ethyl or methyl ether, dioxane, methylene chloride, chloroform, is desirably used to permit better mixing and control of the reaction. The use of greater than 1 :1 molar ratio of arylhalocarbonyl ketene to alcohol or temperatures above 30 C. produces malonic acid diesters. For example, when two moles of alcohol is used per mole of arylhalocarb onyl ketene, the corresponding diester of the arylmalonic acid is produced. Half-amides of arylmalonic acid esters are obtained by reacting the arylhalocarbonyl ketenes with an alcohol followed by reaction of the resulting arylcarboxy ketene ester with a primary or secondary amine as is described herein. Isolation of the intermediate ester is not necessary. A tertiary amine may be used as acid acceptor to remove the hydrogen halide produced during formation of the ester.

The arylcarboxy ketene esters produced as described above are excellent acylating agents particularly suited for the acylation of amines to produce a-carboxyarylacetyl amines. They are especially valuable as agents for the acylation of 6- aminopenicillanic acid.

The acylation of G-aminopenicillanic acid is conducted at a temperature of from about 70 C. to about 50 C. and preferably at a temperature of from about 0 C. to about 30 C. The reaction period is generally from a few minutes up to five hours. A reaction-inert solvent such as ethyl acetate, dioxane, tetrahydrofuran, methyl isobutyl ketone, chloroform or methylene chloride is generally used to facilitate stirring and temperature control. It has been found especially convenient to first form the arylcarboxy ketene ester as described above and to use the reaction mixture, without isolation of the arylcarboxy ketene ester, directly in the amine acylation reaction. In such instances an organic base, i.e., a tertiary amine such as triethylamine or other trialkylamine, preferably a tri(lower alkyl)amine, is used to remove the hydrogen halide produced in formation of the arylcarboxy ketene ester. From a practical standpoint, the 6-aminopenicillanic acid is used as its triethylamine salt. For this reason, methylene chloride is a preferred solvent since the triethylamine salt is readily soluble therein. The sodium or potassium salts of aminopenicillanic acid can also be used but the preferred salt is the triethylamine salt because of its greater solubility in the solvent systems used. An excess of the amine to be acylated can, of course, be used as acid acceptor but is generally avoided, not only for economic reasons but also to prevent possible ammonolysis of the ester group. The reaction is desirably conducted under an atmosphere of nitrogen.

The N-acylation reaction can also be conducted in neutral or alkaline aqueous solution by taking advantage of the slower rate of reaction of the arylcarboxy ketene esters with water at neutral or alkaline pH levels relative to the rate of reaction with the amine group. The reaction is conducted at temperatures ranging from just above the freezing point of the aqueous system to about 50 C. and preferably at from 0 C. to about 20 C. To permit attainment of low temperatures and to facilitate reaction, it is advantageous to employ a mixed solvent system, i.e., water plus a water-miscible reaction-inert organic solvent such as dioxane or tetrahydrofuran. The ketene ester, of course, is desirably used as a solution in the same reaction-inert solvent and is preferably added to the aqueous solution of the -aminopenicillanic acid.

The acylated products are isolated by conventional methods. A typical method, for example, comprises evaporating the reaction mixture to dryness under reduced pressure, dissolving the residue in citrate buffer (pH 5.5) and extracting the product therefrom with chloroform. The chloroform extracts are washed with citrate buffer (pH 5.5), dried with anhydrous sodium sulfate and evaporated to dryness. In another method, which is of value for the isolation of acylation products poorly soluble in methylene chloride, or chloroform, the above method is followed but using n-butanol as extracting solvent in place of chloroform. The product remaining after removal of the n-butanol solvent by evaporation is triturated with ether to produce an amorphous solid.

In still another method, essentially a variation of the above methods, saturated sodium bicarbonate (or potassium bicarbonate) is used in place of the citrate buffer. This method, of course, produces the sodium (or potassium) salt of the acylation product. If necessary to obtain a solid product, the salt is triturated with ether.

In yet another method, the residue remaining after removal of the volatiles from the reaction mixture is taken up in water at pH of from about 2.3 to 2.9, usually about pH 2.7, and the free acid form of the acylation product extracted from the acid solution with chloroform, ether, n-butanol or other suitable solvent. The chloroform, ether or n-butanol extract is then washed with aqueous acid (pl-I 2.3-2.9) and the product recovered by lyophilization or by conversion to a solvent-insoluble salt as by neutralization with an n-butanol solution of sodium or potassium 2-ethyl-hexanoate.

The esters are converted by known methods to the corresponding acids as, for example, by mild acid treatment or enzymatically with an esterase such as liver homogenate.

The novel a-carboxyarylmethylpenicillin esters described herein exhibit in vitro activity against a wide variety of microorganisms, including both gram-positive and gram-negative bacteria. Their useful activity can readily be demonstrated by in vitro tests against various organisms in a brainheart infusion medium by the usual two-fold serial'dilution technique. Additionally, many of the esters of this invention are also useful antibacterial agents in vivo in animals, including man. Especially valuable in this respect are those esters listed in Table II below, which are effective not only via the subcutaneous (SQ) route of administration but also by the oral (PO) route of administration. This property of oral activity is most surprising and unexpected in view of the absence of such activity in the parent a-carboxyarylmethylpenicillins, e. g., a-carboxybenzylpenicillin.

The in vitro activity of the herein described compounds renders them useful for topical application in the form of ointments, creams and the like, or for sterilization purposes, e.g., sick-room utensils.

The valuable products of this invention, especially those of Table II below, are remarkably effective in treating a number of susceptible gram-negative infections in animals, including man. For this purpose, the pure materials or mixtures thereof with other antibiotics can be employed. They may be administered alone or in combination with a pharmaceutical carrier selected on the basis of the chosen route of administration and standard pharmaceutical practice. For example, they may be administered orally in the form of tablets containing such excipients as starch, milk sugar, certain types of clay, etc., or in capsules alone or in admixture with the same or equivalent excipients. They may also be administered orally in the form of elixirs or oral suspensions which may contain flavoring or coloring agents, or be injected parenterally, that is, for example, intramuscularly or subcutaneously. For parenteral administration they are best used in the form of a sterile aqueous solution which may be either aqueous such as water, isotonic saline, isotonic dextrose, Ringers solution, or non-aqueous such as fatty oils of vegetable origin (cotton seed, peanut oil, corn, sesame) and other non-aqueous vehicles which will not interfere with the therapeutic efficiency of the preparation and are non-toxic in the volume or proportion used (glycerol, propylene glycol, sorbitol). Additionally, compositions suitable for extemporaneous preparation of solutions prior to administration may advantageously be made. Such compositions may include liquid diluents, for example, propylene glycol, diethyl carbonate, glycerol, sorbitol, etc.; buffering agents, as well as local anesthetics and inorganic salts to afford desirable pharmacological properties.

The oral and parenteral dosage levels for the herein described compounds are, in general, on the order of up to 200 mg./kg. and mg./kg. of the body weight per day, respectively.

Many of the penicillin ester compounds of this invention, as noted above, exhibit improved absorption on oral administration over that produced by the corresponding free acid or alkali metal salt forms. They, therefore, represent convenient and effective dosage forms of the a-carboxyarylmethylpenicillins of formula 111 above. Of particular interest because of this are the compounds of Table II below.

Further, many of the esters described herein, although inactive or of relatively low activity against Gram-negative organisms per se are, when administered orally to animals, including man, metabolized to the parent acid, i.e., a-carboxybenzylpenicillin, which has a wide spectrum of activity against Grampositive and Gramnegative bacteria. The rate of metabolic conversion of such esters to the parent acid occurs at such a rate as to provide one effective and prolonged concentration of the parent acid in the animal body. In effect, such esters act as depot sources for the parent acid. Especially useful in this respect are those compounds wherein the ester group is COOR wherein R is naphthyl, S-indanyl and 5-(l,2,3,4-

tetrahydronaphthyl).

. under standardized conditions in which nutrient broth containing various concentrations of the test material was seeded with the particular organism specified, and the minimum growth (MIC) at which growth of each organism failed to occur was observed and recorded. The test materials were tested as their sodium or potassium salts. Benzylpenicillin (K salt) when thus tested gave values ofO. 156, 100 vs. S. aureus and E. coli, respectively. (Table 1) TABLE 1 In vitro Data of a-Carboxybenzylpenicillin Esters Against Staphylococcus areus and Escherichia coli MIC mcgJ ml.)

R S. aureus E. culi l-naphthyl 0.39 6.25 S-indanyl 0.39 3.12 1,3-benzodioxolyl) 1.56 6.25 Z-naphth yl 0.78 3.12 G-quinolyl 1.56 50 3-( 2-methyl-4-pyronyl) 0.78 50 H l .25 3. l 2 phenyl" 0.39 6.25 o-methylphenyl 0.05 6.25 m-methylphenyl 0.78 6.25 p-methylphenyl 0.39 6.25 oethylphenyl 0.04 6.25 o-isopropylphenyl 0.39 3.12 o-acetylphenyl' 50 200 p-acetylphenyl 25 200 o-nitrophenyl 25 200 2,3'dimethylphenyl 0.02 3.12 3 ,4dirnethylphenyl 0.04 1.56 2.3-dimethoxyphenyl 1.56 50 2-ch1oro-5-methylpbenyl 0.78 3.12 4-chloro-3-methylphenyl 0.05 6.25 2-chloro-3,4-dimethylphenyl 1.56 25 2-chloro-4;5-dimethylphenyl 3.12 50 4-chioro-2,3-dimethylphenyl 0.01 1.56 4-chloro-2,S-dimethylphenyl 0.39 3.12 4-cliloro2,-dirnethylphenyl 0.04 6.25 2.4-dichloro-6-methylphenyl 0.39 25 2,4-dichloro-3.5-dim ethylphenyl 1.56 25 4-chlor0-2-nitrophenyl 25 200 1-( l .2,3,4-tetrahydronaphthyl) 0.04 6.25 2-( 1 ,4-naphthoquinonyl) 12.5 200 4-indanyl 0.39 6.25 S-quinolyl 1.56 50 6-quinolyl 1.56 50 8-quinolyl 1.56 50 3-( 2-methyl4-pyro'nyl) 0.78 50 2-methoxy-4-( 3-ketobutyl)phenyl 0.7.8 25 4-( 2-carboxyvinyl)phenyl 0.39 50 2-t-butyl-4-methoxyphenyl 0.098 12.5 Z-biphenyl 0.012 1.56 2-carbomethoxymethylphenyl 0.392 6.25 Z-carbethoxymethylphenyl 0.049 6.25 4-( 2-c arboxyethyhphenyl 1.56 50 4-( Z-carbomethoxyethyl)phenyl 0.195 3.125 2-rnethoxy-4( 2- carboxyvinyUphenyl 1.56 25 2-methoxy-4-( 2- carbomethoxyvinyl )phenyl 1.5 6 25 4-( Z-carboxy-Z- carbobenzoxyaminoethyl)phenyl 0.39 100 4-acetamidophenyl 0.78 50 4 -carbomethoxymethylphenyl 1.56 12.5 2-methoxy-4-formylphenyl 0.195 12.5

tested as N-ethylpiperidine salt but also an indicator of the relative rate of hydrolysis of the ester. The lower the MIC value, the greater and more rapid rate of hydrolysis.

Table 11 presents comparative PD values (the dosage which will bring about 50 percent survival of the test animals) for several compounds of this invention in mice (P0 oral and SO subcutaneous routes of administration). The values are obtained under standardized conditions. The procedure comprises production of an acute experimental E. coli 266 infection in mice by the intraperitoneal inoculation of the mice with a standardized (10) E. coli 266 culture suspended in 5 percent hog gastric mucin. The test compounds, in the form of their sodium or potassium salts, are administered to the infected mice by a multiple dosing regimen in which the first dose is given 0.5 hour after inoculation and is repeated 4, 24 and 48 hours later.

The percent of mice surviving after the last treatment are held for four days and the percent survivors then determined. The values thus obtained are then converted to probits and the PD value calculated. The following values are thus obtained (Table 11):

TABLE II Comparative P13 Values of Several Esters of a- Carboxybenzylpenicillin vs. E. coli s-I e) phenyl 66 20 Z-methylphenyl 22- 34 4-methylphenyl 32 48 Z-ethylphenyl 35 50 3-ethylphenyl 25 20 Z-isopropylphenyl 20 l 7 4't-butylphenyl 38 34 4-methoxyphenyl 42 50 Z-ethoxyphenyl 25 37 2.3-dimethylphenyl 29 25 2,4-dimethylphenyl 29 32 2,5-dirnethylphenyl 30 34 2,6-dimethylphenyl 3,4-dirnethylphenyl 21 25 3,5-dimethylphenyl 33 27 2-chlor0-5-methylphenyl 29 29 2-chloro-6-methylphenyl 4 l 27 4-chloro-2-methylphenyl 20 20 4-chloro-3-methylphenyl 43 39 2-methoxy-4-methylphenyl 29 $2 3-methoxy-2-methylphenyl 45 55 4-i'rlethoxy-2-t-butylphenyl 3 2 3 6 4-chloro-2,3-dimethylphenyl 25 1 7 4-chloro -3,S-dimethylphenyl 31 25 4-indanyl 20 27 5-indanyl 38 20 l-tetrahydronaphthyl 25 25 2-tetrahydronaphthyl 42 19 4-(carbomethoxymethyl)phenyl 82 24 Z-biphenyl 16 l 1 1'1 200 30-40 ""potassium salt tested.

*sodium salt tested.

The efficiency of the above esters as pro-drug forms, i.e., substances for biological conversion to the ultimate active principle, for a-carboxybenzylpenicillin is readily evident from the above PD values. The ability of a given ester to protect the test animals against E. coli infection depends upon the rate of absorption of the ester from the gastrointestinal tract and upon the rate of hydrolysis of the ester to a-carboxybenzylpenicillin, the chemotherapeutically-active principle. A ratio of oral PD to subcutaneous PD greater than about 1 to 1 indicates relatively poor absorption of the ester. A high subcutaneous PD value, that is a PD value greater than the subcutaneous PD for a-carboxybenzylpenicillin itself, points to poor hydrolysis of the ester. Most of the above listed esters are equivalent to or better than a-carboxybenzylpenicillin in activity and thus represent convenient, effective oral dosage forms of a-carboxybenzylpenicillin.

The phenyl ester of a-carboxybenzylpenicillin is seen to be about oneythird as effective by the oral route as it is when given subcutaneously due to poor absorption. In other words, more than three times the amount of phenyl ester must be administered orally to achieve the same effect as a given amount of acarboxybenzylpenicillin administered subcutaneously. The 2,6-dimethylphenyl ester is found to be equipotent by the oral and subcutaneous routes of administration. However, despite the favorable ratio of oral to subcutaneous 1"D values, it is less potent than a-carboxybenzylpenicillin and other esters cited above due to poor hydrolysis which gives rise to a relatively high subcutaneous PD Many of the orally active esters of this invention are readily absorbed following oral administration and, even more important from a therapeutic standpoint, are hydrolyzed to the corresponding a-carboxyarylmethylpenicillins. This is evident from the significant serum and urinary levels of the a-carboxyarylmethylpenicillins detected following oral administration of such esters of a-carboxybenzylpenicillin.

Tables 11] and IV below present the mean serum concentrations (as ug. a-carboxybenzylpenicillin) and the mean urinary excretion (as percentage of administered dose) in man following a single oral dose of a-[carbo-(S-indanyloxy)]benzylpenic illin sodium salt or a-[carbo-( 2-isopropylphenoxy)]-benzylpenicillin sodium salt. Similar data for disodium a-carboxybenzylpenicillin administered intramuscularly is presented for comparison purposes in each table. Oral administration of disodium a-carboxybenzylpenicillin to man results in the appearance of no detectable drug in the serum and less than 1 30 percent of the administered dose in the urine (Knudsen et al., Brit. Med. J. 3, 75-8, 1967).

TABLE III Mean Serum Concentrations of oz-Carboxybenzylpenicillin and Certain esters Thereof in Man Serum Concentration oral 10000 8.6 14.6120 7.10.000

"im intramuscular expressed at ug. a-carboxybenzylpenicillin/ml.

The data in Table III demonstrate that oral administration of the two esters and especially of a-[carbo-(Z- isopropylphenoxy)1benzylpenicillin provides serum levels substantially equal to, or superior to, serum levels achieved by intramuscular administration of a-carboxybenzylpenicillin. Oral administration of a-carboxybenzylpenicillin produces, as previously noted, no detectable drug in the serum.

TABLE IV Mean urinary Excretion Levels of a-Carboxybenzylpenicillin and Certain Esters Thereof in Man Urinary Extretionof Administered Dose a-Carboxybenzyl- Hours After Dose Penicillin Route Dose 0-3 3-6 6-24 24-28 5-indanyl ester oral 1000 13.4 15.8 10.3 0

mg. 2-isopropylphenyl 7O ester oral 1000 18.3 15.6 15.6 0

mg. disodium salt im" 500 20.9 12.0 18.5 0

""im intramuscular EXAMPLE 1 Phenylchlorocarbonyl Ketene A. To phenylmalonic acid (20 g.) in ethyl ether (100 ml.) there is added phosphorous pentachloride (46 g.). A vigorous reaction occurs. The reaction mixture is refluxed for 4 hours then the ether partially removed by heating on a steam bath. The reaction mixture becomes black when about half the ether is removed and the remaining ether is removed under reduced pressure (at 100 mm.). The residue is distilled under vacuum and the fraction boiling at 75-90 C. at 1.5-4 mm. collected. The product, a yellow liquid, is redistilled at 74 C. and 1.5 mm. It shows a strong peak in the infrared region of the spectrum at 4.69

Repetition of this procedure but using 10 g. of phenylmalonic acid instead of 20 g. produces a less vigorous reaction on addition of the phosphorous pentachloride. The same product is obtained.

B. Phosphorous pentachloride (23 g.) is added over a fiveminute period to a stirred solution of phenylmalonic acid (10 g.) in ethyl ether (50 ml.) initially at a temperature of 0-5 C. The temperature rises to 13 C. during the addition The mixture is then refluxed for five hours and allowed to stand overnight at room temperature. Removal of the ether at 20 mm.

produces a dark concentrate which is vacuum distilled to give the desired product: b.p. -88 C. at 1.5-2.0 mm. and 74 C. at 0.2 mm.

C. To a stirred solution of phosphorous pentachloride (46 g.) in ethyl ether ml.) there is added phenylmalonic acid (10 g.) over a 2-minute period. The mixture is stirred at room temperature for 4 hours then refluxed for 4 hours and allowed to stand overnight at room temperature. The excess phosphorous pentachloride is filtered off and the ether boiled ofi at atmosphere pressure. The reaction mixture gradually progresses in color from dark yellow to red. The residue is distilled in vacuo to give the product: b.p. 83-86 C. at 1.5 mm. as a yellow liquid.

D. Repetition of this procedure but using an equivalent amount of phosphorous oxychloride as halogenating agent in place of phosphorous pentachloride produces the same product.

EXAMPLE II The procedure of Example l-C is repeated but using the appropriate malonic acid derivative in place of phenylamonic acid to produce the following compounds.

o-methoxyphenyl m-methoxyphenyl p-methoxyphenyl o-trifluoromethylphenyl p-trifluoromethylphenyl m-trifluoromethylphenyl o-isopropylphenyl p-di-( n-propyl )aminophenyl p-chlorophenyl m-chlorophenyl o-butoxyphenyl o-dimethylaminophenyl o-diethylaminophenyl m-dimethylaminophenyl p-dimethylaminophenyl o-dibutylaminophenyl EXAMPLE lll Repetition of the procedures of Examples l-C and 11 but using PBr in place of PC], produces the corresponding bromo compounds.

2-methoxy-4-( 2- carbomethoxyvinyl)-phenyl 2 ,4-dichloro-4-( 2- carboxyvinyl)phenyl 2-methyl-5- (carboxymethyl )phenyl 2dimethylamino-S-biphenyl EXAMPLE IX To a solution of the appropriate aryl halocarbonyl ketene (0.1 mole) in methylene chloride (sufiicient to provide a clear solution and generally from about 5 to ml. per gram of ketene) there is added the proper alcohol R OH (0.1 mole). The reaction mixture is maintained under an atmosphere of nitrogen and stirred for a period of from minutes to three hours, care being taken to exclude moisture. The temperature may range from about 70 C. to about 20 C. The infrared spectrum of the mixture is then taken to determine and confirm the presence of the ketene ester. A solution of 6- aminopenicillanic acid-triethylamine salt (0.1 mole) in methylene-chloride (50 ml.) is added and the mixture stirred at 70 to 20 C. for 10 minutes. The cooling bath is then removed and the reaction mixture stirred continuously and allowed to warm to room temperature. The product is isolated by one of the methods below.

o-ethylphenyl m-ethylphenyl o-n-propylphenyl p-n-propylrhenyl o-isopropy phenyl p-t-butylphenyl o-methoxyphenyl m-methoxyphenyl p-methoxyphenyl o-ethoxyphenyl m-ethoxyphenyl p-ethoxyphenyl p-n-butoxyphenyl o-chlorophenyl m-chlorophenyl p-chlorophenyl o-bromophenyl m-bromophenyl m-fluorophenyl p-fluoro henyl o-formy Fhenyl m-formy phenyl o-acetylphenyl p-acetylphenyl p-butyrylphenyl o-nitrophenyl m-nitrophenyl p-nitrophenyl o-carbomethoxyphenyl m-carbomethoxyphenyl p-earbomethoxyphenyl o-carbethoxyphenyl p-carbo-n-propoxyphenyl Method A The reaction mixture is evaporated to dryness pwmbomflmyloxyphenyl under reduced pressure and the residue taken up in citrate o-dimethylarmnophenyl buffer (pl-l 5.5). The product is extracted from the buffer solu- -gi g p q y I oi-nu y ammop eny tron with chloroform. The chloroform extract IS washed with mdimethYlaminoPhenfl citrate buffer (pH 5.5) then dried with anhydrous sodium m-di-n-propylaminophenyl sulfate and evaporated to dryness to give the sodium salt. p-dimethylammo hen l Method B The procedure of Method A is followed but p-methylethylaminophenyl using n-butanol as extracting solvent in place of chloroform. The product obtained after evaporation of the n-butanol solvent is triturated with ether to give an amorphous solid.

Method C This procedure, a variation of Method A, uses a saturated aqueous solution of sodium (or potassium) bicarbonate in place of citrate buffer to produce the sodium (or potassium) salt of the penicillin product. It is generally used for the recovery of those penicillin products which are poorly soluble in methylene chloride or chloroform.

Method D The reaction mixture is extracted twice with saturated aqueous sodium or potassium bicarbonate, washed with water, dried and evaporated to dryness to give the sodium (or potassium) salt. The product, if not a solid, is triturated with ether.

Method B This method, a modification of Method D, is used for those penicillins which are difiicultly soluble in methylene chloride. The sodium (or potassium) bicarbonate ggjgim ggwflg solution (Method D) is extracted with n-butanol, the butanol 3:5-dimeth l-4 ethyiiahenyl extract dried and evaporated to dryness. g.g-gg yi: tgyipg yi Method F :Thrs method 18 used to isolate the free acid form g' i g ggg 0f the pemclllln- 2,3,4,6-tetramethylphenyl The residue remaining after evaporation of the reaction 3,6-dieth l-2,4-dimeth l henyl mixture to dryness is taken up in aqueous acid, e. g., HCl at pH *fifi'iifii 2,7 and the product extracted therefrom by means of n-bumamin ylpli enyl tanol. The butanol extract is washed with aqueous acid (pH 2,6-dichlorophenyl 2.7) and then lyophilized. gg-gg g l Method F-l The butanol extract of method F is neutral- 2:3:4Lr i2iiigr o3ignyl ized with an n-butanol solution of potassium 2-ethyl hex- 2,4,6-trichlorophenyl anoate to precipitate the potassium salt of the penicillin. g i g' g p y The following compounds are thus prepared. 2:3:5:2: i?a 1i i ri;p lienyl 2,3,4,6-tetrabromophenyd pentachlorophenyl pentabromophenyl 0 2,3,4,6-tetranitrophenyl 2 Method M p ntanitrophenyl 2,4 difluorophenyl 2,6-difluorophenyl phenyl 8 Na 2,3,5-trifluorophenyl o-tolyl B Na 2,4,6-trifluorophenyl m-tolyl B Na 2,3,5,6-tetrafluorophenyl p-tolyl B Na pentafluorophenyl p-methylisopropylaminophenyl 2,3-dimethylphenyl 2,4-dimethylphenyl 2,5-dirnethylphenyl 2,6-dimethylphenyl 3 ,4-dimethylphenyl 3 ,S-dimethylphenyl 2.3-diethylphenyl 2,4-diethylphenyl 2,6-diethylphenyl 2,4-di-n-propylphenyl 2,6-di-n-propylphenyl 5-ethyl-3-methylphenyl 6-ethyl-2-methylphenyl 2-ethyl-4-methylphen l 2-methyl-6-n-propyl henyl 4-isopro yl-3-methy phenyl 2-methy 4-sec-butylphenyl 3-methyl-4-t-butylphenyl 2,3 ,4-trimethylphenyl 2,4.6-trimethylphenyl 2,4-dinitrophenyl 2,6-dinitr0phenyl 3.5-dinitrophenyl 2,3,6-trinitrophenyl OOOOOO UUUUUUUUUUUUUUUUTT T T T T T T OOOOOOOOO WW WWWWW 1 -naphthyl B Na Z-naphthyl B Na l-( 5,6,7,S-tetrahydronaphthyl) B Na 2-( 5,6,7,8-tetrahydronaphthy1) 8 Na 3-( 2-methyl-4-pyronyl) B Na 4-quinolyl F-l K 7-quinolyl F-l K 5-( 1,4-naphthoquinonly) B Na 6-( l,4-naphthoquinonyl) B Na o-(coumarinyl) C K 4-thianaphthenyl B Na l-phenazinyl F-l K Z-phenazinyl F-l K Z-(anthraquinonyl) B Na 4-( o-methylquinolyl) F-l K l-fluorenyl A Na 2-fluorenyl A Na 6-( 1,2-naphthoquinonyl) B Na 7-(1,2-naphthoquinonyl) B Na l-anthraquinonyl B Na 7 -isoquinolyl F-l K 5-( l ,3-benzodioxolyl) K 3-(4-pyronyl) K l-( 3-methyl5,6,7,8-

tetrahydronaphthyl) Na 2-( 4-methyl-5 ,6,7,8-

tetrahydronaphthyl) Na l-(4,8 dimethyl-5 ,6,7.8-

tetrahydron aphthyl) l-( 3.4,6-trimethyl-5,6,7,8-

tetrahydronaphthyl) 2-( 3,4,6-trimethyl-5,6,7,8-

tetrahydronaphthyl) 2-( 5,5,8,8-tetramethyl5,6,7,8-

tetrahydronaphthyl) 2-acetyl-S-dimethylaminophenyl 2-acetyl-5-ethoxyphenyl 2-butyryl-3,S-dimethylphenyl Z-furyl 3-furyl 3-( N-methyl)pyridyl 4-( N-methyl)pyridyl Z-biphenyl 3-biphenyl 2-chloro-3-biphenyl 2-acetyl-4-biphenyl Z-dimethylamino-5-biphenyl 2-t-butyl-4-methoxyphenyl 4-carbethoxymethylphenyl 4-( carbo n-butoxy)methylphenyl 4-( Z-carbethoxyethyDethyl 2-methoxy-4-(2- carbomethoxyvinyl)phenyl 4-( Z-carbomethoxyvinyl)phenyl Z-hydroxymethylphenyl 2-isopropyl-4-methoxyphenyl 3-dimethylamino-4 isopropylphenyl 2-t-butyl-4-isopropylphenyl 4( 2-carbomethoxyethyl )phenyl 2-chloro-3-( Z-carboxyethyl )phenyl 2-methoxy-4-formylphenyl EXAMPLE Xlll The sodium and potassium salts of Examples Xl-Xll are converted to the corresponding acids by careful neutralization of aqueous solutions of their salts with aqueous phosphoric acid followed by extraction of the acid form into methylisobutylketone. The methylisobutylketone solutions are washed with water, dried with anhydrous sodium sulfate, filtered and evaporated to give the free acids.

EXAMPLE XlV The free acids of Examples lX-Xlll are converted to their calcium, magnesium, ammonium, procaine, N,N-

dibenzylethylenediamine, N-ethylpiperidine, dibenzylamine, l-ephenamine, triethylamine, N-benzyl-B-phenethylamine, N,N'-bis(dehydroabietyl)ethylenediamine and benzhydrylamine salts by reaction of aqueous solutions thereof with one equivalent of the appropriate base. The salts are recovered by freeze drying.

EXAMPLE XV A solution of the triethylamine salt of a-[carbo(5-indanyloXy)]-benzy1 penicillin (0.5 g.) in a small volume of saturated gram and the presence of a-carboxybenzylpenicillin along with a small amount of benzyl penicillin.

EXAMPLE XVI -A solution of the sodium salt of a-[carbo(5-indanyloxy)]- benzyl penicillin in water (0.5 g. in 5 ml.) is held at room temperature for 24 hours. The pH is automatically regulated at 8.0-8.5 by the addition of sodium bicarbonate. The solution is then freeze dried and the byproduct phenol removed by trimration of the residue with ethanol to give the disodium salt.

Repetition of this procedure but at 35 C. for 2 hours also produces the disodium salt.

EXAMPLE XVll A tablet base is prepared by blending the following ingredients in the proportion by weight indicated.

Sucrose, U.S.P. 8 Tapioca starch 1 magnesium stearate pus noui Sufiicient a-[carbo( S-indanyloxy) ]benzyl penicillin triethylamine salt is blended into the base to provide tablets containing 25', and 250 mg. of active ingredient.

EXAMPLE XVlll Capsules containing 25, 100 and 250 mg. of active ingredient are prepared by blending sufiicient a-[carbo(5-in danyloxy)]benzyl penicillin sodium salt inthe following mixture (proportions given by parts in weight).

Calcium carbonate. U.S.P. 17.6 Dicalcium phosphate 18.8 Magnesium trisilicate 5.2 Lactose. U.S.P. 5.2 Potato starch 5.2 Magnesium stearate A 0.8 Magnesium stearate B 0.35

EXAMPLE XIX A suspension of a-[carbo(4-chloro-2,3-dimethylphenoxy) ]benz yl penicillin potassium salt is prepared with the following composition:

Penicillin compound g- 31.42 70% Aqueous sorbitol g--7l4.29 Glycerine, U.S.P. g--l85.35 Gum acacia (10% solution) ml-l00 Polyvinyl pyrrolidone g-- 0.5 Propyl parahydroxybenzoate g- 0.172 Distilled water to make one liter g- 0.094

Various sweetening and flavoring agents may be.added to this suspension, as well as acceptable colors. The suspension contains approximately 25 mg. of penicillin compound per milliliter.

EXAMPLE XX The sodium salt of a-[carbo-(o-isopropylphenoxy)]benzyl penicillin (10 g.) is intimately mixed and ground with sodium citrate (4% by weight). The ground, dry mixture is filled into vials, sterilized with ethylene oxide and the vials sterilely stoppered. For parenteral administration, sufficient water is added to the vials to form solutions containing 25 mg. of active ingredient per ml.

PREPARATION A Malonic Acids The following aryl malonic acids not previously described in the literature are prepared by the method of Wallingford et al., J. Am. Chem. Soc. 63, 2056-2059 (i964) which comprises condensing an alkyl carbonate, usually diethyl carbonate, with an equimolar proportion of the desired ethyl aryl acetate in the presence of an excess (4-8 times) of sodium ethylate with continuous removal of by-product alcohol from the reaction mixture. The esters thus produced are hydrolyzed to the acid by known methods.

C O OH d-Cg C O OH R, RI

3-furyl 3-pyridyl o-methoxyphenyl 4-pyridyl o-butoxyphenyl o-dimethylaminophenyl o-diethylaminophcnyl m-dimethylaminophenyl p-dirnethylaminophenyl o-dibutylaminophenyl m-methoxyphenyl p-methoxyphenyl o-trifluoromethylphenyl m-trifluoromethylphenyl p-trifluoromethylphenyl o-isopropylphenyl p-di-( n-propyl)aminophenyl The necessary o-trifluoromethylphenylacetic acid is prepared from otrifluoromethylbenzonitrile by the procedure of Corse, et al., J. Am Chem. Soc. 70, 2841 (1948) which comprises: (a) conversion of the nitrile to o-trifluoromethylacetophenone by a Grignard reaction with methylmagnesium iodide followed by hydrolysis; (b) reaction of the acetophenone with sulfur and morpholine at l35 C. for 16 hours followed by treatment with glacial acetic acid and hydrochloric acid.

What is claimed is: l. A compound of the formula:

l kyl)amino and trifluoromethyl;

andR is wherein Z is lower alkylene and is selected from the group consisting of (CH and (CH and substituted derivatives thereof wherein the substituent is selected from the group consisting of methyl, chloro and bromo.

2. The compounds of claim 1 wherein R is phenyl and the pharmaceutical]y-acceptable salts thereof.

3. The compounds of claim 1 wherein R is chlorophenyl and the pharmaceutically-acceptable salts thereof. 30 4. The compound of claim 2 wherein Z is (CH;.);, and

the pharmaceuticalldv-acceptable salts thereof.

5. The compoun of claim 3 wherein R 1S o-chlorophenyl and R is 2-(5,6,7,8-tetrahydronaphthyl) and the pharmaceutically-acceptable salts thereof. I

6. a-[carbo( 5-indanyloxy)]benzylpenicillin.

7. An antibacterial composition comprising a pharmaceutis cally-acceptable carrier and an antibacterial amountof a compound according to claim 1.

8. A method of controlling bacteria which comprises con- 40 tacting an area infected with said bacteria with an antibacterial amount of a compound according to claim 1.

UNITED STATES PATENT orrrca CERTHHCATE @F QQRREE'HQN Patent NO. 9, 01 t d y 1972 lnventor(s) n Kenneth Butler It is certified that error appears in the above-identified patent and that said Letters Patent are hereby corrected as shown below:

Col l line 31, after "Org. Syntheses 20," should appear Vol. 47 line 57, Spheuyl should read 8-phenyl Col. 3, line 67, "(CH should read [CH Col 6, line 70, "one"should read an Col 7, line 16, "areus" should read aureus line 53, "2-carbethoxymethylphenyl" should read 4-carbethoxymethylphenyl Col 9 line 2, "oneythird" should read one-third Table III "Hours after Dose"should appear under "Serum Concentration." Table III, line 40, numbers "4 6 8 24" should appear over the I appropriate columns. Table III, line 42, the first occurrence of "10.2" should be omitted. Table III lines 43, 46 G 48, "mg." should appear after the dose levels in each case. Col ll, line 64, "2-diphenyl" should read 2-biphnyl line 69, "4 (carb0" should read 4- (carbo- Col 12, line 22, "2,4dinitrophenyl" should read 2,4-dinitrophenyl Col 13, line 57, "2,7" should read 2.7 Col 14, line 64,. "2,3,4,6-tetrabromophenyd" should read 2,3,4,6-tetra bromophenyl 1 Col 18, line 42, Pmethylethylaminophenyl" should read p-methylethylaminophenyl 7 line 53, "4-butyl452-methylphenyl" should read 4-butyl-5iso-.

propyl-Z-methylphenyl Col 19, line 36, "b" should read B under "Method" column.

line 53, A should appear under column "Method. Col. 22, line 45, "6tbutyl" should read 6tbutyl Col 24, line 37, "4acetyl2 fluoro-S-nitrophenyl" should appear under R column. line 49 "Na" should appear under column "M." Col 28, claim 1, that portion of the formula reading "N-CH-OOH" should read N-CH -COOH Signed and sealed this 30th day of January 1973 L a (SEAL) Attest:

EDWARD MDFLETCHER, JR. I ROBERT GOTTSCHALK Commissioner of Patents Attesting Officer 

2. The compounds of claim 1 wherein R1 is phenyl and the pharmaceutically-acceptable salts thereof.
 3. The compounds of claim 1 wherein R1 is chlorophenyl and the pharmaceutically-acceptable salts thereof.
 4. The compound of claim 2 wherein Z'' is -(CH2)3- and the pharmaceutically-acceptable salts thereof.
 5. The compound of claim 3 wherein R1 is o-chlorophenyl and R2 is 2-(5,6,7,8-tetrahydronaphthyl) and the pharmaceutically-acceptable salts thereof.
 6. Alpha -(carbo(5-indanyloxy))benzylpenicillin.
 7. An antibacterial composition comprising a pharmaceutically-acceptable carrier and an antibacterial amount of a compound according to claim
 1. 8. A method of controlling bacteria which comprises contacting an area infected with said bacteria with an antibacterial amount of a compound according to claim
 1. 